Abstract 2369: The NRF2 and dimeric STAT3 complex coordinately regulates <i>IL-23</i> expression: Implications for breast cancer progression
Authors: Su-Jung Kim, Soma Saeidi, Nam-Chul Cho, Seung Hyeon Kim, Han-Byoel Lee, Wonshik Han, Hye-Kyung Na, Young-Joon Surh
Journal: Cancer Research (2022)

Abstract Basal-like breast cancer (BLBC) is an aggressive malignancy with a poor prognosis. Persistent activation of NRF2 and STAT3 is considered to stimulate the aggressive behavior of BLBC. An online database, NRF2-ome predicted the possibility of interplay between NRF2 and STAT3 based on domain motif interactions. However, cooperation between NRF2 and STAT3 and its implications for breast cancer progression remain elusive. Immunoprecipitation and in situ proximity analyses revealed that NRF2 and STAT3 directly interact with each other, which predominantly occurred in the nucleus of human breast cancer cells. After confirming the physical interaction between STAT3 and NRF2, we examined whether this could influence the expression or stability of each transcription factor. While silencing of NRF2 failed to alter the STAT3 expression, STAT3 knockdown facilitated degradation of NRF2. Of note, silencing of STAT3 did not affect the mRNA expression of NRF2, suggesting that STAT3-induced accumulation of NRF2 is attributable to stabilization of the preexisting protein. As the NRF2-STAT3 complex was found to accumulate predominantly in the nucleus, we speculate that NRF2 could bind to a functionally active phosphorylated form of STAT3 (P-STAT3Y705). The substitution of Y705 of STAT3 with the non-phosphorylatable phenylalanine (STAT3Y705F) abolished interaction with NRF2. Notably, the dimeric form, but not a monomeric form, of STAT3 interacts directly with the Neh1 and Neh3 domains of NRF2. Next, we investigated the functional significance of STAT3- NRF2 interaction in BLBC growth in vivo. STAT3 knockdown significantly reduced expression of NRF2 in the xenograft tumor tissues derived from MDA-MB-231 human breast cancer cells. Tumors derived from cells transfected with the non-specific control siRNA showed a pronounced interaction between STAT3 and NRF2, whereas such interaction was abolished in the STAT3-silenced xenograft tumor. Analysis of the TCGA multi-omics data showed that high levels of STAT3 and NRF2 mRNA transcripts were correlated with elevated expression of P-STAT3Y705 and NRF2 target proteins in breast cancer patients. Next generation RNA sequencing analysis identified the gene encoding interleukin-23A (IL-23A) upregulated by concurrent binding of STAT3 and NRF2 to its promoter. IL-23A is markedly overexpressed in BLBC, compared with other subtypes of breast cancer. Combined silencing of NRF2 and STAT3 inhibited BLBC growth to a greater extent than that achieved with single siRNA transfection. IL-23A depletion by siRNA also showed the similar phenotypic changes to those caused by double knockdown of both transcription factors. In conclusion, these findings suggest that the STAT3-NRF2 interaction accelerates BLBC growth and progression by augmenting IL-23A expression, which underscores the importance of subtype-specific molecular pathways in human breast cancer. Citation Format: Su-Jung Kim, Soma Saeidi, Nam-Chul Cho, Seung Hyeon Kim, Han-Byoel Lee, Wonshik Han, Hye-Kyung Na, Young-Joon Surh. The NRF2 and dimeric STAT3 complex coordinately regulates IL-23 expression: Implications for breast cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2369.